Tumor therapeutic agent

ABSTRACT

A novel tumor therapeutic agent for a combination therapy is disclosed. More specifically, a tumor therapeutic agent used for a combination therapy of lenvatinib, ifosfamide, and etoposide is disclosed.

TECHNICAL FIELD

The present invention relates to a tumor therapeutic agent used for acombination therapy of lenvatinib, ifosfamide, and etoposide.

BACKGROUND ART

Lenvatinib has angiogenesis inhibitory effect (Patent Literature 1) andinhibitory effect (Patent Literatures 2 to 5) against tyrosine kinase,which is reported to be involved in the malignant transformation oftumors (Non-Patent Literatures 1 to 5), and lenvatinib is known as atherapeutic agent for various tumors, such as thyroid cancer, lungcancer, melanoma, endometrial cancer, gastric cancer, bladder cancer,renal cancer, glioma, liver cancer, and ovarian cancer.

Ifosfamide is known as a therapeutic agent for many malignant tumors,such as small cell lung cancer, prostate cancer, cervical cancer, andosteosarcoma.

Etoposide is known as a therapeutic agent for malignant tumors, such assmall cell lung cancer, malignant lymphoma, and acute leukemia.

In general, tumor therapeutic agents are often not effective for allpatients when they are used alone. Accordingly, attempts have been madeso far to improve the treatment rate by using a plurality of tumortherapeutic agents in combination to enhance antitumor effects andreduce adverse effects (Patent Literature 6 and Patent Literature 7).

The combined administration of ifosfamide and etoposide is used formalignant tumors, such as osteosarcoma and Ewing's sarcoma (Non-PatentLiteratures 6 to 8).

CITATION LIST Patent Literature

-   Patent Literature 1: U.S. Patent Application Publication No.    2004-053908-   Patent Literature 2: U.S. Patent Application Publication No.    2004-253205-   Patent Literature 3: U.S. Patent Application Publication No.    2010-105031-   Patent Literature 4: U.S. Patent Application Publication No.    2009-209580-   Patent Literature 5: U.S. Patent Application Publication No.    2009-264464-   Patent Literature 6: U.S. Patent Application Publication No.    2011-104161-   Patent Literature 7: U.S. Patent Application Publication No.    2004-259834

Non Patent Literature

-   Non-Patent Literature 1: Lasota et al., “Mutations in Exons 9 and 13    of KIT Gene Are Rare Events in Gastrointestinal Stromal Tumors,”    American Journal of Pathology, vol. 157, pp. 1091-1095, 2000-   Non-Patent Literature 2: Berdel et al., “Recombinant Human Stem Cell    Factor Stimulates Growth of a Human Glioblastoma Cell Line    Expressing c-kit Protooncogene1,” Cancer Research, vol. 52, pp.    3498-3502, 1992-   Non-Patent Literature 3: Lennartsson et al., “The stem cell factor    receptor/c-Kit as a drug target in cancer,” Current Cancer Drug    Targets, vol. 6, pp. 65-75, 2006-   Non-Patent Literature 4: Turner et al., “Fibroblast growth factor    signalling: from development to cancer,” Nature Reviews Cancer, vol.    10, pp. 116-129, 2010-   Non-Patent Literature 5: Wells et al, “Targeting the RET Pathway in    Thyroid Cancer,” Clinical Cancer Research, vol. 15, pp. 7119-7123,    2009-   Non-Patent Literature 6: The ESMO/European Sarcoma Network Working    Group, “Bone sarcomas: ESMO Clinical Practice Guideline for    diagnosis, treatment and follow-up,”Annals of Oncology, vol. 23,    supplement 7, vii100-vii109, 2012-   Non-Patent Literature 7: Gentet et al., “Ifosfamide and etoposide in    childhood osteosarcoma. A phase II study of the French Society of    Paediatric Oncology,” European Journal of Cancer, vol. 33, pp.    232-237, 1997-   Non-Patent Literature 8: Grier et al., “Addition of Ifosfamide and    Etoposide to Standard Chemotherapy for Ewing's Sarcoma and Primitive    Neuroectodennal Tumor of Bone,” The New England Journal of Medicine,    vol. 348, pp. 694-701, 2003

SUMMARY OF INVENTION Technical Problem

However, the therapeutic effects of the combined use of tumortherapeutic agents ever reported are not sufficient, and development ofcombination therapy using further new tumor therapeutic agents has beenawaited.

Solution to Problem

In view of the above circumstances, the present inventors conductedextensive research, and consequently found that combined administrationof lenvatinib, ifosfamide, and etoposide surprisingly showed antitumoreffects more than expected; thus, the present invention has beencompleted.

That is, the present invention provides the followings:

-   [1] A therapeutic agent for a tumor comprising lenvatinib or a    pharmacologically acceptable salt thereof, wherein lenvatinib or the    pharmacologically acceptable salt thereof is administered in    combination with ifosfamide and etoposide or a pharmacologically    acceptable salt thereof.-   [2] A pharmaceutical composition comprising lenvatinib or a    phannacologically acceptable salt thereof, wherein lenvatinib or the    pharmacologically acceptable salt thereof is administered in    combination with ifosfamide and etoposide or a pharmacologically    acceptable salt thereof.-   [3] A pharmaceutical composition for treating a tumor, comprising    lenvatinib or a pharmacologically acceptable salt thereof; wherein    lenvatinib or the pharmacologically acceptable salt thereof is    administered in combination with ifosfamide and etoposide or a    pharmacologically acceptable salt thereof.-   [4] A method for treating a tumor, comprising administering    lenvatinib or a pharmacologically acceptable salt thereof,    ifosfamide, and etoposide or a pharmacologically acceptable salt    thereof to a patient in need thereof.-   [5] Lenvatinib or a pharmacologically acceptable salt thereof,    wherein the lenvatinib or a pharmacologically acceptable salt    thereof is administered in combination with ifosfamide and etoposide    or a pharmacologically acceptable salt thereof.-   [6] Lenvatinib or a pharmacologically acceptable salt thereof for    treating a tumor, wherein the lenvatinib or a pharmacologically    acceptable salt thereof is administered in combination with    ifosfamide and etoposide or a pharmacologically acceptable salt    thereof.-   [7] A pharmaceutical composition comprising lenvatinib or a    pharmacologically acceptable salt thereof for treating a tumor,    wherein lenvatinib or the pharmacologically acceptable salt thereof    is administered in combination with ifosfamide and etoposide or a    pharmacologically acceptable salt thereof.-   [8] Use of lenvatinib or a pharmacologically acceptable salt thereof    for the manufacture of a pharmaceutical composition for treating a    tumor, wherein lenvatinib or the pharmacologically acceptable salt    thereof is administered in combination with ifosfamide and etoposide    or a pharmacologically acceptable salt thereof.-   [9] Use of lenvatinib or a pharmacologically acceptable salt thereof    for treating a tumor, wherein lenvatinib or the pharmacologically    acceptable salt thereof is administered in combination with    ifosfamide and etoposide or a pharmacologically acceptable salt    thereof.-   [10] A kit comprising a formulation comprising lenvatinib or a    pharmacologically acceptable salt thereof; a formulation comprising    ifosfamide, and a formulation comprising etoposide or a    pharmacologically acceptable salt thereof.-   [11] The kit according to [10], wherein the kit is a kit for    treating a tumor.-   [12] A therapeutic agent for a tumor comprising lenvatinib or a    pharmacologically acceptable salt thereof; ifosfamide, and etoposide    or a pharmacologically acceptable salt thereof.-   [13] A pharmaceutical composition comprising lenvatinib or a    pharmacologically acceptable salt thereof, ifosfamide, and etoposide    or a pharmacologically acceptable salt thereof.-   [14] A pharmaceutical composition for treating a tumor, the    pharmaceutical composition comprising lenvatinib or a    pharmacologically acceptable salt thereof, ifosfamide, and etoposide    or a pharmacologically acceptable salt thereof.-   [15] Lenvatinib or a pharmacologically acceptable salt thereof,    ifosfamide, and etoposide or a pharmacologically acceptable salt    thereof for treating a tumor.-   [16] Use of lenvatinib or a pharmacologically acceptable salt    thereof, ifosfamide, and etoposide for the manufacture of a    therapeutic agent for a tumor.-   [17] The above therapeutic agent for treating a tumor or    pharmaceutical composition, further comprising an excipient.-   [18] The above therapeutic agent for treating tumor, pharmaceutical    composition, treatment method, use, compound, or kit, wherein the    lenvatinib or a pharmacologically acceptable salt thereof is    lenvatinib mesylate.-   [19] The above therapeutic agent for treating a tumor,    pharmaceutical composition, treatment method, use, compound, or kit,    wherein the etoposide or a pharmacologically acceptable salt thereof    is etoposide phosphate.-   [20] The above therapeutic agent for treating tumor, pharmaceutical    composition, treatment method, use, compound, or kit, wherein the    lenvatinib or a pharmacologically acceptable salt thereof,    ifosfamide, and etoposide or a pharmacologically acceptable salt    thereof are administered simultaneously, separately, continuously,    or at time intervals.-   [21] The above therapeutic agent for treating a tumor,    pharmaceutical composition, treatment method, use, compound, or kit,    wherein the tumor is osteosarcoma, Ewing's sarcoma,    rhabdomyosarcoma, neuroblastoma, retinoblastoma, hepatoblastoma, or    nephroblastoma.-   [22] The therapeutic agent for treating a tumor, pharmaceutical    composition, treatment method, use, compound, or kit according to    [21], wherein the tumor is osteosarcoma.-   [23] The therapeutic agent for treating a tumor, pharmaceutical    composition, treatment method, use, compound, or kit according to    [22], wherein the osteosarcoma is relapsed or refractory.

Advantageous Effects of Invention

The present invention provides a therapeutic agent for a tumor used fora combination therapy of lenvatinib, ifosfamide, and etoposide. Thistherapeutic agent for a tumor exhibits unexpected antitumor effects forpatients in need thereof.

DESCRIPTION OF EMBODIMENTS

Embodiments of the present invention are described below. The followingembodiments are only examples for describing the present invention, andit is not intended to limit the present invention only to theseembodiments. The present invention can be carried out in various formswithout departing from the subject matter of the present invention.

Note that the documents, publications, patent publications, and otherpatent literatures cited in the present description are incorporatedherein by reference. The present specification includes the contents ofU.S. Provisional Application No. 62/207,546 (filed on Aug. 20, 2015),which is the basis for claiming the priority of the present application.

Lenvatinib refers to4-(3-chloro-4-(cyclopropylaminocarbonyl)aminophenoxy)-7-methoxy-6-quinolinecarboxamide,and the structural formula thereof is shown in the following formula:

Lenvatinib or a pharmacologically acceptable salt thereof can beproduced by the method described in Patent Literature 1. An example ofpharmacologically acceptable salts of lenvatinib is lenvatinib mesylate.Lenvatinib mesylate is also referred to as E7080.

Ifosfamide refers to3-(2-chloroethyl)-2[(2-chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine-2-oxide,and the structural formula thereof is shown in the following formula:

Ifosfamide can be produced by a known method, and can be produced, forexample, by the method described in U.S. Pat. No. 3,732,340. Moreover,ifosfamide can be obtained by purchasing Ifex (registered trademark)from Bristol-Myers Squibb, or by purchasing Ifomide (registeredtrademark) from Shionogi & Co., Ltd.

Etoposide refers to(5R,5aR,8aR,9S)-9-{[4,6-O-(1R)-ethylidene-β-D-glucopyranosyl]oxy}-5-(4-hydroxy-3,5-dimethoxyphenyl)-5,8,8a,9-tetrahydrofuro[3′,4′:6,7]naphtho[2,3-d][1,3]dioxol-6(5aH)-one,and the structural formula thereof is shown in the following formula:

Etoposide can be produced by a known method, and can be produced, forexample, by the method described in U.S. Pat. No. 3,524,844. Moreover,etoposide can be obtained by purchasing VePesid (registered trademark)from Bristol-Myers Squibb. An example of pharmacologically acceptablesalts of etoposide is etoposide phosphate.

Although the “pharmacologically acceptable salt” is not limited to aspecific type of salt, examples thereof include salts with inorganicacids, salts with organic acids, salts with inorganic bases, salts withorganic bases, and salts with acidic or basic amino acids.

Examples of salts with inorganic acids include salts with hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid.Examples of salts with organic acids include salts with acetic acid,succinic acid, fumaric acid, maleic acid, tartaric acid, citric acid,lactic acid, stearic acid, benzoic acid, methanesulfonic acid (mesylicacid), ethanesulfonic acid, and p-toluenesulfonic acid.

Examples of salts with inorganic bases include alkali metal salts, suchas sodium salt and potassium salt; alkaline earth metal salts, such as acalcium salt and a magnesium salt; an aluminum salt and an ammoniumsalt. Examples of salts with organic bases include salts withdiethylamine, diethanolamine, meglumine, andN,N-dibenzylethylenediamine.

Examples of salts with acidic amino acids include salts with asparticacid and glutamic acid. Examples of salts with basic amino acids includesalts with arginine, lysine, and ornithine.

Examples of pharmacologically acceptable salts of lenvatinib includesalts with organic acids, and one embodiment thereof is methanesulfonate(mesylate).

Examples of pharmacologically acceptable salts of etoposide includesalts with inorganic acids, and one embodiment thereof is phosphate.

When solvates and optical isomers of the lenvatinib, ifosfamide, oretoposide of the present invention are present, these solvates andoptical isomers are included. Examples of solvates include hydrate andanhydrate. Examples of solvents include water, alcohols (e.g., methanol,ethanol, and n-propanol), and dimethylformamide.

Moreover, the lenvatinib, ifosfamide, or etoposide of the presentinvention may be crystalline or amorphous. When polymorphic crystals arepresent, a single crystal form or a mixture of any of these crystalforms may be used.

The dose of lenvatinib or a pharmacologically acceptable salt thereofcan be suitably selected, depending on the degree of symptoms, thedevelopment of adverse effects, the age, sex, body weight, andsensitivity difference of the patient, administration route,administration period, administration interval, the type ofpharmaceutical formulation, etc.

The dose of lenvatinib or a pharmacologically acceptable salt thereof isnot particularly limited, but is generally, when orally administered toan adult (body weight: 60 kg) or an child, 0.1 to 500 mg, 0.5 to 300 mg,or 1 to 100 mg, per day, or is 0.1 to 500 mg/m² (body surface area,hereinafter the same), 0.5 to 300 mg/m², or 1.0 to 100 mg/², per day.This dose can be generally administered once a day, or two to threetimes a day. If the patient has undergone excessive toxicity, it isnecessary to reduce the dose. The dose and dosage schedule may bechanged when one or more additional chemotherapeutic agents are used, inaddition to the combination therapy of the present invention. The dosageschedule can be determined by the physician who is treating the specificpatient.

Ifosfamide can be administered according to known clinical practice. Thedose and dosage schedule can be changed according to the specificdisease symptoms and the total symptoms of the patient The dose can besuitably reduced, depending on age, sex, symptoms, development ofadverse effects, etc.

The dose of ifosfamide is not particularly limited, but is generally,when administered to an adult (body weight: 60 kg) or an child orally,intravenously, or by intravenous drip infusion, 0.01 to 100 g/m², 0.05to 30 g/m², or 0.1 to 10 g/m², per day, or 0.01 to 100 g, 0.05 to 30 g,or 0.1 to 10 g per day. One embodiment of the dose of ifosfamide isabout 1.5 to 3 g/m² per day. This dose can be generally administered forfirst 3 to 5 days of one cycle consisting of 3 to 4 weeks. Ifosfamidecan be repeatedly administered to the patient by repeating this cycle.In one embodiment, about 3 g/m² per day of ifosfamide is administeredfrom Day 1 to Day 3 of one cycle consisting of 21 days, and this cycleis suitably repeated. If the patient has undergone excessive toxicity,it is necessary to reduce the dose. The dose and dosage schedule may bechanged when one or more additional chemotherapeutic agents are used.The dosage schedule can be determined by the physician who is treatingthe specific patient.

Etoposide or a pharmacologically acceptable salt thereof can beadministered according to known clinical practice. The dose and dosageschedule can be changed according to the specific disease symptoms andthe total symptoms of the patient. The dose can be suitably reduced,depending on age, sex, symptoms, development of adverse effects, etc.

The dose of etoposide or a pharmacologically acceptable salt thereof isnot particularly limited, but is generally, when administered to anadult (body weight: 60 kg) or an child orally, intravenously, or byintravenous drip infusion, 1.0 to 5000 mg/m², 5.0 to 3000 mg/m², or 10to 1000 mg/m², per day. One embodiment of the dose of etoposide or apharmacologically acceptable salt thereof is about 60 to about 120 mg/m²as etoposide per day. This dose can be generally administered for first3 to 5 days of one cycle consisting of 3 to 4 weeks. Etoposide or apharmacologically acceptable salt thereof can be repeatedly administeredto the patient by repeating this cycle. If the patient has undergoneexcessive toxicity, it is necessary to reduce the dose. In oneembodiment, about 100 mg/m², as etoposide per day, of etoposide or apharmacologically acceptable salt thereof is administered from Day 1 toDay 3 of one cycle consisting of 21 days, and this cycle is suitablyrepeated.

The dose and dosage schedule may be changed when one or more additionalchemotherapeutic agents are used. The dosage schedule can be determinedby the physician who is treating the specific patient.

The dose of lenvatinib, ifosfamide, or etoposide in the combinedadminishation of the present invention can generally be set equal to orlower than the dose when they are administered alone. The specific dose,administration route, administration frequency, administration cycle,etc., are suitably determined, in consideration of the age, sex, andsymptoms of the patient, the development of adverse effects, etc.

The dosage form of lenvatinib, ifosfamide, and etoposide in the presentinvention is not particularly limited, and it is only necessary toadminister lenvatinib, ifosfamide, and etoposide in combination whenadministered. For example, lenvatinib, ifosfamide, and etoposide areeach administered to the patient simultaneously, separately,continuously, or at time intervals. Here, the term “simultaneously”means that the ingredients are each administered in the same period orexactly at the same time, or administered through the same route. Theterm “separately” means that the ingredients are each administered indifferent intervals or with different frequencies, or administeredthrough different routes. The term “continuously” means that theingredients are each administered through the same or different routesin an arbitrary order within a predetermined period. The term “at timeintervals” means that the ingredients are each administered through thesame or different routes at intervals for the respective ingredients. Inthe combined administration of lenvatinib and ifosfamide, whenlenvatinib is administered during one cycle of administration ofifosfamide described above or during a period in which this cycle isrepeated, it is determined that lenvatinib and ifosfamide areadministered in combination. In the combined administration oflenvatinib and etoposide or a pharmacologically acceptable salt thereof,when lenvatinib is administered during one cycle of administration ofetoposide or a pharmacologically acceptable salt thereof described aboveor during a period in which this cycle is repeated, it is determinedthat lenvatinib and etoposide or a pharmacologically acceptable saltthereof are administered in combination. In one dosage form in thecombined administration of the present invention, lenvatinib is orallyadministered, and ifosfamide and etoposide are administeredintravenously or by intravenous infusion. Moreover, the combinedadministration of the present invention may be conducted simultaneously,separately, continuously, or at time intervals, together with theadministration of a therapeutic agent for a tumor other than lenvatinib,ifosfamide, and etoposide.

The therapeutic agent for a tumor of the present invention can beformulated, for example, by the method described in the 16th RevisedJapanese Pharmacopoeia (JP), United States Pharmacopeia (USP), orEuropean Pharmacopoeia (EP).

The tumor therapeutic agent of the present invention can be orallyadministered in the form of a solid formulation, such as a tablet, agranule, a fine granule, a powder, or a capsule; or a liquidformulation, a jelly, or a syrup.

Moreover, the tumor therapeutic agent of the present invention may beparenterally administered in the form of an injection, a suppository, anointment, a cataplasm, or the like.

When an oral solid formulation is prepared, an excipient, and further abinder, a disintegrator, a lubricant, a coloring agent, a flavoringagent, a surfactant, etc., can be added, if necessary, as additives tolenvatinib or a pharmacologically acceptable salt thereof, ifosfamide,or etoposide or a pharmacologically acceptable salt thereof, therebyforming a tablet, a granule, a subtle granule, a powder, a capsule,etc., by a conventional method. The above additives can be suitablycombined for formulation. Furthermore, these tablets, granules, etc.,may be coated, if necessary

When an injection is prepared, a pH adjuster, a buffer, a suspendingagent, a solubilizing agent, a stabilizer, an isotonizing agent, apreservative, etc., may be added, if necessary, to lenvatinib or apharmacologically acceptable salt thereof, ifosfamide, or etoposide or apharmacologically acceptable salt thereof, thereby forming anintravenous, subcutaneous, or intramuscular injection, or an intravenousinfusion by a conventional method. In this case, the injections can beformed, if necessary, as lyophilized products by a conventional method.

Tumors targeted by the present invention are not particularly limited,and examples of tumors include brain tumors (including hypophysialadenoma and glioma), head and neck cancer, cervical cancer, jaw cancer,maxillary cancer, submandibular gland cancer, oral cancer (includingtongue cancer, mouth floor cancer, gingival cancer, buccal mucosacancer, and hard palate cancer), salivary gland cancer, sublingual glandcancer, parotid gland cancer, nasal cavity cancer, paranasal sinuscancer (including maxillary sinus cancer, frontal sinus cancer, ethmoidsinus cancer, and sphenoid sinus cancer), laryngeal cancer (includingsupraglottic cancer, glottic cancer, and subglottic cancer), esophagealcancer, lung cancer (including bronchogenic cancer, non-small cell lungcancer (including lung adenocarcinoma, squamous cell carcinoma, andlarge cell lung carcinoma), small cell lung cancer (including oat cell(lymphoid) and intermediate cell types), and mixed small cell/large celllung cancer), breast cancer, pancreatic cancer (including pancreaticduct cancer), gastric cancer (including scirrhous gastric cancer andundifferentiated gastric cancer (including undifferentiatedadenocarcinoma, signet-ring cell carcinoma, and mucinous carcinoma)),biliary tract cancer (including bile duct cancer and gallbladdercancer), small intestinal or duodenal cancer, colorectal cancer(including colon cancer, rectal cancer, cecal cancer, sigmoid coloncancer, ascending colon cancer, transverse colon cancer, and descendingcolon cancer), bladder cancer, renal cancer (including renal cellcancer), hepatic cancer (including liver cell cancer and intrahepaticbile duct cancer), prostate cancer, uterine cancer (including cervicalcancer and endometrial cancer), ovarian cancer, thyroid cancer,pharyngeal cancer (including nasopharyngeal cancer, oropharyngealcancer, and hypopharyngeal cancer), sarcomas (e.g., osteosarcoma,Ewing's sarcoma, rhabdomyosarcoma, chondrosarcoma, Kaposi's sarcoma,angiosarcoma, fibrosarcoma, etc.), neuroblastoma, retinoblastoma,hepatoblastoma, nephroblastoma, malignant lymphomas (including Hodgkin'slymphoma and non-Hodgkin's lymphoma), leukemia (including, for example,chronic myelocytic leukemia (CML), acute myelocytic leukemia (AML),chronic lymphocytic leukemia (CLL) and acute lymphocytic leukemia (ALL),lymphoma, multiple myeloma (MM), myelodysplastic syndrome, etc.), skincancer (including basal cell cancer, squamous cell cancer, malignantmelanoma, mycosis fungoides, Sezary's syndrome, solar keratosis, Bowen'sdisease, and Paget's disease) and melanoma; and mixed tumors thereof. Inone embodiment of the present invention, the target tumor isosteosarcoma, Ewing's sarcoma, rhabdomyosarcoma, neuroblastoma,retinoblastoma, hepatoblastoma, or nephroblastoma. In another embodimentof the present invention, the target tumor is osteosarcoma. Moreover,the tumors targeted by the present invention include relapsed orrefractory tumors.

EXAMPLES

Specific examples of the present invention are provided below; however,the present invention is not limited thereto.

Example 1

Tumor growth inhibitory activity of the combined use of lenvatinib,ifosfamide, and etoposide against human osteosarcoma cell line (143B)subcutaneous transplantation models

Using 6 NOD SCID mice (NOD.CB17-Prkdc^(scid)/J, female mice, CharlesRiver Laboratories Japan, Inc.) in each group, antitumor effects wereevaluated when lenvatinib alone, a combination of two agents, i.e.,ifosfamide and etoposide, or a combination of three agents, i.e.,lenvatinib, ifosfamide, and etoposide, was administered. Cells ofosteosarcoma cell line 143B (ATCC CRL-8303) derived from human weresuspended at a concentration of 1×10⁷ cells/mL, and 0.1 mL of thesuspension was subcutaneously transplanted to the right flank of eachmouse.

The longest diameter and short axis of the tumor were measured with anelectronic digital caliper (Digimatic (tread mark) Caliper, MitutoyoCorporation) 9 days after transplantation. The mice were divided intogroups so that the average value of the tumor volume of each group wasalmost equal (Day 1). The tumor volume was calculated according to thefollowing equation:

Tumor volume=longest diameter(mm)×short axis(mm)×short axis(mm)/2

Lenvatinib mesylate was dissolved in purified water to a concentrationof 0.3 mg/mL. Ifosfamide (Sigma Aldrich) was dissolved in aphysiological saline solution to a concentration of 25 mg/mL. Moreover,an etoposide injection (20 mg/mL solution, Bristol-Myers Squibb) wasdiluted with a physiological saline solution to a concentration of 0.8mg/mL.

Purified water and a physiological saline solution were administered toa control group. A lenvatinib solution and a physiological salinesolution were administered to a lenvatinib group. An ifosfamidesolution, an etoposide solution, and purified water were administered toan ifosfamide+etoposide combination group. A lenvatinib solution, anifosfamide solution, and an etoposide solution were administered to a3-agent combination group.

The administration dose of lenvatinib mesylate was 3 mg/kg, and 10 mL/kgwas orally administered once a day for 7 days (Day 1 to Day 7). Theadministration dose of ifosfamide was 250 mg/kg, and 10 mL/kg wasintraperitoneally administered once (Day 1). The administration dose ofetoposide was 8 mg/kg, and 10 mL/kg was intraperitoneally administeredonce a day for 3 days (Day 2 to Day 4). Purified water was orallyadministered in an amount of 10 mL/kg once a day for 7 days (Day 1 toDay 7). A physiological saline solution was intraperitoneallyadministered in an amount of 10 mL/kg once a day for 4 days (Day 1 toDay 4).

The tumor volume of each mouse was measured 8 days (Day 8) after thestart of administration. The results are shown in Table 1. It wasclarified that the combined use of the three agents, i.e., lenvatinib,ifosfamide, and etoposide, had excellent antitumor effects against thehuman osteosarcoma cell-line (143B) subcutaneous transplantation models.

TABLE 1 Average tumor volume Group (Day 8) mm³ Control group 785.4Lenvatinib group 438.6 Ifosfamide + etoposide combination group 495.93-agent combination group 184.7

Example 2

Tumor growth inhibitory activity of the combined use of lenvatinib,ifosfamide, and etoposide against human osteosarcoma cell line (G-292clone A141B1) subcutaneous transplantation models

Using 6 NOD SCID mice (NOD.CB17-Prkdc^(scid)/J, female mice, CharlesRiver Laboratories Japan, Inc.) in each group, antitumor effects wereevaluated when lenvatinib alone, a combination of two agents, i.e.,ifosfamide and etoposide, or a combination of three agents, i.e.,lenvatinib, ifosfamide, and etoposide, was administered. Cells ofosteosarcoma cell line G-292 clone A141B1 (JCRB Cell Bank, IFO50107)derived from human were suspended at a concentration of 1×10⁸ cells/mL,and 0.1 mL of the suspension was subcutaneously transplanted to theright flank of each mouse.

The longest diameter and short axis of the tumor were measured with anelectronic digital caliper (Digimatic (tread mark) Caliper, MitutoyoCorporation) 29 days after transplantation. The mice were divided intogroups so that the average value of the tumor volume of each group wasalmost equal (Day 1). The tumor volume was calculated according to thefollowing equation:

Tumor volume=longest diameter(mm)×short axis(mm)×short axis(mm)/2

Lenvatinib mesylate was dissolved in purified water to a concentrationof 0.3 mg/mL. lfosfamide (Sigma Aldrich) was dissolved in aphysiological saline solution to a concentration of 25 mg/mL. Moreover,an etoposide injection (20 mg/mL solution, Bristol-Myers Squibb) wasdiluted with a physiological saline solution to a concentration of 0.8mg/mL.

Purified water and a physiological saline solution were administered toa control group. A lenvatinib solution and a physiological salinesolution were administered to a lenvatinib group. An ifosfamidesolution, an etoposide solution, and purified water were administered toan ifosfamide+etoposide combination group. A lenvatinib solution, anifosfamide solution, and an etoposide solution were administered to a3-agent combination group.

The administration dose of lenvatinib mesylate was 3 mg/kg, and 10 mL/kgwas orally administered once a day for 7 days (Day 1 to Day 7). Theadministration dose of ifosfamide was 250 mg/kg, and 10 mL/kg wasintraperitoneally administered once (Day 1). The administration dose ofetoposide was 8 mg/kg, and 10 mL/kg was intraperitoneally administeredonce a day for 3 days (Day 2 to Day 4). Purified water was orallyadministered in an amount of 10 mL/kg once a day for 7 days (Day 1 toDay 7). A physiological saline solution was intraperitoneallyadministered in an amount of 10 mL/kg once a day for 4 days (Day 1 toDay 4).

The tumor volume of each mouse was measured 8 days (Day 8) after thestart of administration. The results are shown in Table 2. It wasclarified that the combined use of the three agents, i.e., lenvatinib,ifosfamide, and etoposide, had excellent antitumor effects against thehuman osteosarcoma cell line (G-292 clone A141B1) subcutaneoustransplantation models.

TABLE 2 Average tumor volume Group (Day 8) mm³ Control group 362.5Lenvatinib group 267.9 Ifosfamide + etoposide combination group 286.03-agent combination group 208.9

Example 3

Tumor growth inhibitory activity of the combined use of lenvatinib,ifosfamide, and etoposide against human osteosarcoma cell line (HOS)subcutaneous transplantation models

Using 6 NOD SOD mice (NOD.CB17-Prkdc^(scid)/J, female mice, CharlesRiver Laboratories Japan, Inc.) in each group, antitumor effects wereevaluated when lenvatinib alone, a combination of two agents, i.e.,ifosfamide and etoposide, or a combination of three agents, i.e.,lenvatinib, ifosfamide, and etoposide, was administered. Cells ofosteosarcoma cell line HOS (Human Science Research Resources Bank,IFO50106) derived from human were suspended at a concentration of 1×10⁸cells/mL, and 0.1 mL of the suspension was subcutaneously transplantedto the right of each mouse.

The longest diameter and short axis of the tumor were measured with anelectronic digital caliper (Digimatic (tread mark) Caliper, MitutoyoCorporation) 18 days after transplantation. The mice were divided intogroups so that the average value of the tumor volume of each group wasalmost equal (Day 1). The tumor volume was calculated according to thefollowing equation:

Tumor volume=longest diameter(mm)×short axis(mm)×short axis(mm)/2

Lenvatinib mesylate was dissolved in purified water to a concentrationof 0.3 mg/mL. Ifosfamide (Sigma Aldrich) was dissolved in aphysiological saline solution to a concentration of 25 mg/mL. Moreover,an etoposide injection (20 mg/mL solution, Bristol-Myers Squibb) wasdiluted with a physiological saline solution to a concentration of 0.8mg/mL.

Purified water and a physiological saline solution were administered toa control group. A lenvatinib solution and a physiological salinesolution were administered to a lenvatinib group. An ifosfamidesolution, an etoposide solution, and purified water were administered toan ifosfamide+etoposide combination group. A lenvatinib solution, anifosfamide solution, and an etoposide solution were administered to a3-agent combination group.

The administration dose of lenvatinib mesylate was 3 mg/kg, and 10 mL/kgwas orally administered once a day for 7 days (Day 1 to Day 7). Theadministration dose of ifosfamide was 250 mg/kg, and 10 mL/kg wasintraperitoneally administered once (Day 1). The administration dose ofetoposide was 8 mg/kg, and 10 mL/kg was intraperitoneally administeredonce a day for 3 days (Day 2 to Day 4). Purified water was orallyadministered in an amount of 10 mL/kg once a day for 7 days (Day 1 toDay 7). A physiological saline solution was intraperitoneallyadministered in an amount of 10 mL/kg once a day for 4 days (Day 1 toDay 4).

The tumor volume of each mouse was measured 8 days (Day 8) after thestart of administration. The results are shown in Table 3. It wasclarified that the combined use of the three agents, i.e., lenvatinib,ifosfamide, and etoposide, had excellent antitumor effects against thehuman osteosarcoma cell line (HOS) subcutaneous transplantation models.

TABLE 3 Average tumor volume Group (Day 8) mm³ Control group 468.6Lenvatinib group 352.2 Ifosfamide + etoposide combination group 433.03-agent combination group 252.1

Example 4

Phase 1/2 clinical trial of lenvatinib for children and adolescents withrelapsed or refractory solid malignant tumors

The present clinical trial is a phase 1/2 clinical trial for evaluatingthe safety, tolerance, and efficacy of lenvatinib in children andadolescents who have relapsed or refractory solid malignant tumors.

The present clinical trial consists of 5 cohorts (Cohorts 1, 2A, 2B, 3A,and 3B), and a primary object of each cohort is as follows:

To determine the recommended dose (RD) of lenvatinib in children andadolescents who have relapsed or refractory solid malignant tumors usingthe time-to-event continual reassessment method (TiTE-CRM) [Cohort 1].In the present clinical trial, the RD is defined as a dose at which theincidence of dose limiting toxicity (DLT) is closest to the targetlevel, i.e., 20%.

To evaluate the objective response rate (ORR) of lenvatinib in childrenand adolescents who have 131 iodine-refractory differentiated thyroidcancer (DTC) [Cohort 2A]. The objective response rate is defined ascomplete response (CR)+partial response (PR), based on the ResponseEvaluation Criteria in Solid Tumors (RECIST 1.1).

To evaluate the progression-free survival (PFS) at 4 months afteradministration of lenvatinib alone in children and adolescents who haverelapsed or refractory osteosarcoma [Cohort 2B]. The progression-freesurvival (PFS) at 4 months is the rate of participants who survive for 4months after the start of administration of lenvatinib alone and whosedisease does not progress (based on RECIST 1.1).

To set the RD of lenvatinib when used in combination with ifosfamide andetoposide in children and adolescents who have relapsed or refractorysolid malignant tumors [Cohort 3A]. The dose of a combination oflenvatinib, ifosfamide, and etoposide at which DLT occurs in one or lessof the 6 cases is taken into consideration as the RD of combinationtherapy.

To evaluate the progression-free survival (PFS) at 4 months afteradministration of a combination of lenvatinib, ifosfamide, and etoposidein children and adolescents who have relapsed or refractory osteosarcoma[Cohort 3B]. The progression-free survival (PFS) at 4 months is the rateof participants who survive for 4 months after the start of combined useof lenvatinib, ifosfamide, and etoposide and whose disease does notprogress (based on RECIST 1.1).

Secondary objects of the present clinical trial are as follows:

Best overall response (BOR) [all cohorts]

Objective response rate (ORR) [Cohorts 1, 2B, 3A, and 3B]

Duration of response (DOR) [all cohorts]

Disease control rate (DCR) [all cohorts]

Clinical benefit rate (CBR) [all cohorts]

Time to progression (TTP) [all cohorts]

Overall survival (OS) [all cohorts]

The number of participants who experienced adverse events (AEs)/seriousadverse events (SAEs) as the safety and tolerance evaluation

Plasma concentration of lenvatinib: area under the drugconcentration-time curve (AUC) [all cohorts]

Pharmacodynamics (PD) biomarkers: VEGFR and FGFR [all cohorts]

The present clinical trial consists of 5 cohorts,

In Cohort 1 (single agent dose setting), in order to set the recommendeddose (RD) of lenvatinib in children and adolescents who have relapsed orrefractory solid malignant tumors, the dose is gradually increased usingthe time-to-event continual reassessment method (TiTE-CRM). Once the RDis set, Cohorts 2A, 2B, and 3A are started in parallel.

In Cohort 2 (single agent expansion cohort), the efficacy of lenvatinibat the RD in children and adolescents who have 131 iodine-refractorydifferentiated thyroid cancer [Cohort 2A] or relapsed or refractoryosteosarcoma [Cohort 2B] is evaluated.

In Cohort 3A (combined-use dose setting), the RD of lenvatinib when usedin combination with ifosfamide and etoposide in participants who haverelapsed or refractory osteosarcoma is determined.

In Cohort 3B (combined-use expansion cohort), the efficacy of thecombined use of lenvatinib, ifosfamide, and etoposide at the RD obtainedin Cohort 3A in participants who have relapsed or refractoryosteosarcoma is evaluated. Participants with osteosarcoma who areregistered in Cohort 1 or 2B and whose disease have progressed afteradministration of lenvatinib become candidates for participants ofCohort 3B, as with participants with relapsed or refractory osteosarcomato which lenvatinib has not been administered.

In the present clinical trial, lenvatinib is provided as a hard capsulecontaining 1.225 mg, 4.9 mg, or 12.25 mg of lenvatinib mesylate (1 mg, 4mg, or 10 mg as lenvatinib). For children who cannot swallow capsules,the lenvatinib capsule is dissolved in water.

In Cohort 1, lenvatinib is orally administered once a day at a startingdose of 11 mg/m² from Day 1 to Day 28 of a 28-day cycle. The dose can bereduced to 9 mg/m², or increased to 14 and 17 mg/m².

In Cohort 2A, lenvatinib is orally administered once a day at the RDdetemined in Cohort 1 from Day 1 to Day 28 of a 28-day cycle.

In Cohort 2B, lenvatinib is orally administered once a day at the RDdetermined in Cohort 1 from Day 1 to Day 28 of a 28-day cycle.

In Cohort 3A, lenvatinib is orally administered once a day at a dose(starting dose) 20% lower than the RD determined in Cohort 1 from Day 1to Day 21 of a 21-day cycle. Ifosfamide is administered at a dose of3000 mg/m²/day (starting dose) from Day 1 to Day 3 of a 21-day cycle for5 cycles in total. Etoposide is administered at a dose of 100 mg/m²/day(starting dose) from Day 1 to Day 3 of a 21-day cycle for 5 cycles intotal.

In Cohort 3B, lenvatinib is orally administered once a day at the RDdetermined in Cohort 3A from Day 1 to Day 21 of a 21-day cycle.Ifosfamide is administered at the dose set in Cohort 3A from Day 1 toDay 3 of a 21-day cycle for 5 cycles in total. Etoposide is administeredat the dose set in Cohort 3A from Day 1 to Day 3 of a 21-day cycle for 5cycles in total.

From the results of the present clinical study, the efficacy, safety,and tolerance of lenvatinib alone for 131 iodine-refiactorydifferentiated thyroid cancer, those of lenvatinib alone for relapsed orrefractory osteosarcoma, and those of the 3-agent combined use oflenvatinib, ifosfamide, and etoposide for relapsed or refractoryosteosarcoma can be confirmed.

1. A pharmaceutical composition for treating a tumor, comprisinglenvatinib or a pharmacologically acceptable salt thereof, whereinlenvatinib or the pharmacologically acceptable salt thereof isadministered in combination with ifosfamide and etoposide or apharmacologically acceptable salt thereof.
 2. The pharmaceuticalcomposition according to claim 1, further comprising an excipient. 3.The pharmaceutical composition according to claim 1, wherein thelenvatinib or a pharmacologically acceptable salt thereof is lenvatinibmesylate.
 4. The pharmaceutical composition according to claim 1,wherein the etoposide or a pharmacologically acceptable salt thereof isetoposide phosphate.
 5. The pharmaceutical composition according toclaim 1, wherein the lenvatinib or a pharmacologically acceptable saltthereof, ifosfamide, and etoposide or a pharmacologically acceptablesalt thereof are administered simultaneously, separately, continuously,or at time intervals.
 6. The pharmaceutical composition according toclaim 1, wherein the tumor is osteosarcoma, Ewing's sarcoma,rhabdomyosarcoma, neuroblastoma, retinoblastoma, hepatoblastoma, ornephroblastoma.
 7. The pharmaceutical composition according to claim 6,wherein the tumor is osteosarcoma.
 8. The pharmaceutical compositionaccording to claim 7, wherein the osteosarcoma is relapsed orrefractory.
 9. (canceled)
 10. A method for treating a tumor, comprisingadministering lenvatinib or a pharmacologically acceptable salt thereof,ifosfamide, and etoposide or a pharmacologically acceptable salt thereofto a patient in need thereof.
 11. (canceled)
 12. The method according toclaim 10, wherein the lenvatinib or a pharmacologically acceptable saltthereof is lenvatinib mesylate.
 13. The method according to claim 10,wherein the etoposide or a pharmacologically acceptable salt thereof isetoposide phosphate.
 14. The method according to claim 10, wherein thelenvatinib or a pharmacologically acceptable salt thereof, ifosfamide,and etoposide or a pharmacologically acceptable salt thereof areadministered simultaneously, separately, continuously, or at timeintervals.
 15. The method according to claim 10, wherein the tumor isosteosarcoma, Ewing's sarcoma, rhabdomyosarcoma, neuroblastoma,retinoblastoma, hepatoblastoma, or nephroblastoma.
 16. The methodaccording to claim 15, wherein the tumor is osteosarcoma.
 17. The methodaccording to claim 16, wherein the osteosarcoma is relapsed orrefractory.